This invention relates generally to an apparatus for separation of liquid compounds by column chromatography. More particularly, the present invention relates to a miniaturized chromatography column for the rapid separation of organic chemicals, inorganic chemicals, biochemicals and biomolecules by liquid phase chromatography.
It has been found that traditional column chromatography apparatus are not easily adapted for the rapid separation of biomolecules in a laboratory setting. Where there is a need for separation of compounds from a body fluid, such as blood or urine, a laboratory technician must set up individual columns for each type of separation to be done. The process of setting up a column consists of selecting the appropriate stationary phase medium specific to the biochemical or body fluids to be separated and packing the column with the appropriate stationary phase medium. After setting up the conventional column chromatography apparatus, the technician is then able to introduce the eluent into the column and collect the eluate, then recover the separated biochemical from the stationary phase media by appropriate washings.
Conventional column chromatography apparatus provide inherently slow separations due to the length of the column or the quantity of biochemical to be separated. Numerous methods of accelerating the separations have been developed, such as use of a pump to forcedly move the eluent through the column or plungers to maintain a pressure gradient on the eluent. None of the conventional column chromatography apparatus, however, are well adapted for diagnostic, analytical or other laboratory procedures where rapid separations of biochemicals or biomolecules from body fluids are required.
Conventional column chromatography apparatus consist of glass buret-like tubes which are packed with a separatory medium. The size and length of these conventional apparatus make them unsuitable for use in rapid separations with high recovery yields. Further, conventional chromatography apparatus require the repetitive transfer of the eluate or subsequent washes to a multitude of vessels. The necessity for the use of many vessels to collect the eluate and wash increases the likelihood of contamination in the separated biochemical and, therefore, increases the likelihood of erroneous analytical or diagnostic results. Finally, conventional chromatography apparatus are typically capable of single-stage separations whereby only one type of eluent or one type of separation media may be employed. It has been found desirable to provide a miniaturized column chromatography apparatus which will meet the foregoing deficiencies commonly found with conventional column chromatography apparatus. The present invention, therefore, is directed to providing a column chromatography apparatus which is readily adaptable for rapid laboratory separation of biochemicals and biomolecules from an eluent fluid, such as a body fluid. In particular, it has been determined that a miniaturized column chromatography apparatus having a multi-stage separatory tube, an associated eluent tube, a vented-cap and a non-vented cap wherein the entire apparatus is sized to be usable with a common laboratory centrifuge or, if necessary, an ultracentrifuge and is adapted to fit conventional test tubes, holder racks, mini-vials, syringes and other laboratory equipment. By providing a column chromatography apparatus configured in this manner, a laboratory technician is able to rapidly separate biomolecules from the eluent fluid, recover and quantify the desired biochemical.